Effects of trypan blue on thyroid secretion. Inhibition of purified cathepsin D from bovine thyroid.

Previous studies have demonstrated that trypan blue directly inhibits thyroid secretion when the dye is administered in vitro or in vivo. To further study the mechanisms of inhibition, cathepsin D (EC 3.4.23.5) (thyroidal acid proteinase) has been purified from bovine thyroid. Trypan blue inhibited the proteolysis of both 125I-labeled thyroglobulin and 125I-labeled hemoglobin in both crude lysosomal enzyme preparation and purified endopeptidase and the inhibition was competitive. Inhibition was also observed when the dye was allowed to prebind to either purified enzyme or purified substrate. Inhibition of cathepsin D is shown to account for part of the inhibition of thyroid secretion.

Use of osmium-ferrocyanide treatment for improved lysosomal acid trimetaphosphatase reaction and subcellular detail in thyroid follicular cells.

The histochemical reaction for acid trimetaphosphatase in addition to secondary tissue treatment with an osmium-ferrocyanide mixture was used to study lysosomes and phagolysosomes in the mouse thyroid gland. The osmium-ferrocyanide postfixation enhanced reaction product localization, reduced diffuse reaction, and improved membrane contrast. In addition, the ultrathin tissue sections did not require heavy metal staining, thus eliminating potential stain artifacts due to precipitation. In view of the improved tissue preservation and enzyme localization, it is suggested that osmium-ferrocyanide postfixation be used after the acid trimetaphosphatase method.

A psychiatric mother-baby day hospital for pregnant and postpartum women.

Major depression and other psychiatric disorders are common during pregnancy and the postpartum period, yet these disorders remain largely under-diagnosed and under-treated. Developing programs that are uniquely tailored to meet the needs of perinatal psychiatric patients can improve both the quality and acceptability of care. In this report, we describe the development and implementation of a novel mother-baby day hospital service designed to meet the mental health needs of this special population, and present preliminary data regarding treatment acceptability and effectiveness. Our experience using this model of care for the past five years has suggested that specialized units such as this one represent an acceptable, effective, fiscally viable approach to the care of pregnant and postpartum psychiatric patients. Further research is needed to more thoroughly assess the effectiveness of this type of specialized perinatal service.

Patient choice of treatment for postpartum depression: a pilot study.

OBJECTIVE: The lack of systematic efficacy research makes the selection of optimal treatment for postpartum depression (PPD) difficult. Moreover, the treatment decisions for women with PPD who are breastfeeding are heavily influenced by their concerns about infant exposure to antidepressant medication. The objective of this pilot trial was to examine the clinical characteristics of women with PPD associated with treatment selection. METHOD: This open pilot trial offered 23 women with PPD one of 3 treatment options: sertraline, interpersonal psychotherapy (IPT), or their combination administered in an outpatient mental health setting over 12 weeks. Baseline and treatment outcome measures included the Hamilton Rating Scale for Depression (HRSD), the Beck Depression Inventory (BDI) and the Edinburgh Postnatal Depression Scale (EPDS). RESULTS: Completers across all 3 treatment groups (n = 18) experienced significant clinical improvement with each of the 3 treatment modalities on the HRSD (p < 0.001), BDI (p < 0.001) and EPDS (p < 0.001). There were trends for women with a prior depression to more frequently choose sertraline as a treatment (alone or with IPT, p = 0.07), and for women who were breastfeeding to choose sertraline (alone or with IPT, p = 0.10) less frequently. CONCLUSION: In this small sample of women with PPD, most women chose IPT with or without sertraline. A larger randomized study could further confirm the suggested predictors of treatment selection identified in this study: previous depression and breastfeeding status.

Effects of trypan blue on thyroid secretion: localization of trypan blue within the colloid space and phagolysosomes of thyroid follicles.

Trypan blue was previously shown to directly inhibit thyroid secretion following TSH stimulation. Inhibition of both colloid droplet formation and thyroglobulin proteolysis was demonstrated. By observing the characteristic bright red fluorescence of the dye-protein complex, we have demonstrated that trypan blue rapidly enters the colloid space and combines with thyroglobulin. In addition, the dye in association with thyroglobulin has been demonstrated within phagosomes and phagolysosomes by centrifugation of the lysosomal (P15) fraction on both sucrose and Percoll density gradients. Lability or latency of the dye with the phagolysosomal contents was demonstrated and the dye was found in association with thyroglobulin by column chromatography. It is proposed that the complexing of trypan blue to thyroglobulin alters its attachment to specific follicular cell receptors, inhibits pinocytosis, and, thus, thyroid hormone secretion.

Ultrastructural and cytochemical effects of trypan blue on TSH stimulation of thyroid follicular cells.

Ultrastructural and cytochemical techniques were used to study the effects of trypan blue on the response of mouse-thyroid cells to exogenous stimulation by thyroid stimulating hormone (TSH). The dye delayed the response to TSH resulting in decreased colloid-droplet formation in the apical region of the cells. The dye did not stop the shift of trimetaphosphatase activity from lysosomes to phagolysosomes. The duration of the TSH-induced response was shorter in the dye treated thyroids. Small vesicles, with trimetaphosphatase reaction product, were found near Golgi elements, phagolysosomes, and the plasma membrane facing rhe intercellular space of adjacent follicle cells. Their enzyme activity was not affected by exposure to the dye. These data indicate that the primary effect of trypan blue on the response of thyroid follicle cells to TSH stimulation was reduced endocytosis in the apical region resulting in fewer colloid droplets.

Direct effects of trypan blue on thyroid secretion.

Trypan blue directly inhibited in vitro thyroid secretion (butanol soluble 125I release to the media) induced by both thyroid stimulating hormone (TSH) and dibutyryl cAMP. Intracellular colloid droplet counts were also decreased. Inhibition was directly proportional to dye concentration and could be overcome by supramaximal TSH and dibutyryl cAMP. Inhibition could be observed as early as 20 min of incubation, was not increased by preincubation, and could even be demonstrated after TSH in vivo. Trypan blue, in vivo, produced similar inhibition of thyroid secretion. Incubation of 125I-thyroglobulin with lysosomal enzymes revealed inhibition with much lower concentrations of dye. Inhibition of lysosomal enzyme(s) would not appear to explain the marked decreases in colloid droplets, and this may represent two separate effects of trypan blue on thyroid secretion.